Inter- and intraspecific variation in the distribution and number of pit organs (free neuromasts) of sharks and rays

The distribution of pit organs (free neuromasts) has previously been documented for several species of pelagic sharks, but is relatively poorly known for rays and bottom-dwelling (demersal) sharks. In the present study, the complete distribution of pit organs was mapped in the demersal sharks Heterodontus portusjacksoni, Orectolobus maculatus, Hemiscyllium ocellatum, Chiloscyllium punctatum, and Asymbolus analis, and the rays Rhinobatos typus, Aptychotrema rostrata, Trygonorrhina sp. A, Raja sp. A, and Myliobatis australis. All of these species had pit organs scattered over the dorsolateral surface. The sharks also had "mandibular" pit organs (and "umbilical" pit organs in C. punctatum and A. analis) on the ventral surface, while pit organs were sparse or absent on the ventral surface of rays. All of the species examined here, except for M. australis, also had a "spiracular" group of pit organs adjacent to the eye and/or spiracle. Spiracular pit organs were also recorded for the sawshark Pristiophorus sp. A and the skate Pavoraja nitida, although the remainder of pit organs were not mapped in these species. The distribution and number of pit organs varied both within and among species. Pit organ distribution was asymmetrical in each individual examined, but no particular trend towards left or right "handedness" was observed in any species. Although rays have been thought to have fewer pit organs than sharks in general, this was not the case in the present study. All of the species examined here had few pit organs compared to the pelagic sharks previously documented, but it is not clear whether this is due to ecological or phylogenetic causes.     

Fate of the methyl group during the conversion of sterigmatocystin into O-methylsterigmatocystin and aflatoxin B1 by cell-free preparations of Aspergillus parasiticus

Cell-free extracts of fungal mycelia of two aflatoxin non-producing isolates of Aspergillus parasiticus (SRRC 163 and SRRC 2043) were utilized for the study of enzyme activities involved in the latter stages of aflatoxin biosynthesis. The post-microsomal fractions (105,000 x g supernatant) of both SRRC 163 and SRRC 2043 were able to convert sterigmatocystin (ST) into O-methylsterigmatocystin (OMST); whereas the microsomal (105,000 x g pellet) preparation of only SRRC 163 was able to convert OMST into aflatoxin B1 (AFB1). S-Adenosylmethionine (SAM) was the primary substrate for the ST to OMST (methyltransferase) enzymatic conversion; [3H]OMST of specific activity 0.93 Ci/mmol was obtained in a reaction containing the [3H]SAM substrate (specific activity 1 Ci/mmol). After the terminal enzymatic conversion of OMST into AFB1, none of the radiolabel of the methyl group from OMST was found in AFB1. It is postulated that the methylation of ST may be required for subsequent enzymatic oxidation of OMST to aflatoxin B1.     

CUC1 gene activates the expression of SAM-related genes to induce adventitious shoot formation

CUP-SHAPED COTYLEDON (CUC)1 encodes members of the NAC family. These are functionally redundant genes that are involved in shoot apical meristem (SAM) formation and cotyledon separation during embryogenesis in Arabidopsis. We analyzed transgenic plants overexpressing CUC1 (35S::CUC1). The cotyledons of these transgenic seedlings regularly had two basal lobes, small and round epidermal cells between the sinuses, and adventitious SAMs on the adaxial surface of this region. This suggests that CUC1 promotes adventitious SAM formation by maintaining epidermal cells in an undifferentiated state. In 35S::CUC1 cotyledons, the class I knotted-like homeobox (KNOX) genes, including SHOOT MERISTEMLESS (STM) and BREVIPEDICELLUS (BP), which are involved in SAM formation and/or maintenance, were ectopically expressed before adventitious SAM formation. In stm mutants, ectopic expression of CUC1 could not induce adventitious SAMs, whereas they continued to be observed in bp mutants. These results suggest that STM, but not BP, is necessary for the formation of adventitious SAMs in 35S::CUC1 cotyledons. Furthermore, we examined the relationship between CUC1 and ASYMMETRIC LEAVES (AS)1 and AS2. The as1 and as2 mutations genetically enhance 35S::CUC1 phenotypes even in the absence of STM function. Interestingly, the as1 mutation can partially rescue the mutant vegetative development phenotypes in the cuc1 cuc2 double mutant. Our results suggest that CUC1 positively regulates SAM formation not only through STM but also through an STM-independent pathway that is negatively regulated by AS1 and AS2.     

Evaluating the Effectiveness of Self-Administration of Medication (SAM) Schemes in the Hospital Setting: A Systematic Review of the Literature

Background

Self-administration of medicines is believed to increase patients'' understanding about their medication and to promote their independence and autonomy in the hospital setting. The effect of inpatient self-administration of medication (SAM) schemes on patients, staff and institutions is currently unclear.

Objective

To systematically review the literature relating to the effect of SAM schemes on the following outcomes: patient knowledge, patient compliance/medication errors, success in self-administration, patient satisfaction, staff satisfaction, staff workload, and costs.

Design

Keyword and text word searches of online databases were performed between January and March 2013. Included articles described and evaluated inpatient SAM schemes. Case studies and anecdotal studies were excluded.

Results

43 papers were included for final analysis. Due to the heterogeneity of results and unclear findings it was not possible to perform a quantitative synthesis of results. Participation in SAM schemes often led to increased knowledge about drugs and drug regimens, but not side effects. However, the effect of SAM schemes on patient compliance/medication errors was inconclusive. Patients and staff were highly satisfied with their involvement in SAM schemes.

Conclusions

SAM schemes appear to provide some benefits (e.g. increased patient knowledge), but their effect on other outcomes (e.g. compliance) is unclear. Few studies of high methodological quality using validated outcome measures exist. Inconsistencies in both measuring and reporting outcomes across studies make it challenging to compare results and draw substantive conclusions about the effectiveness of SAM schemes.     

Probing fundamental film parameters of immobilized enzymes--towards enhanced biosensor performance. Part I--QCM-D mass and rheological measurements

Enzyme immobilization is an ever-growing research-area for both analytical and industrial applications. Of critical importance in this area are the effects of immobilization procedures upon the functionality of the immobilized biomolecules. Both beneficial and detrimental effects can be conferred through the selection and tuning of the immobilization procedure. Quartz-crystal microbalance with dissipation (QCM-D) has been previously used to great effect in tracking alterations to thin films of biomolecules immobilized onto quartz transducers.In this study, we investigate the ability of QCM-D to track and monitor film parameters of a monolayer of laccase immobilized on a series of self-assembled monolayers (SAMs), differing in lateral density of binding residues on the SAM and height of the SAM from the quartz surface. Both mass gains and rheological parameters for these varying surfaces were measured and trends later compared to the apparent enzyme kinetics of the immobilized laccase films, assessed electroanalytically (Paper II in this two part study). For covalent attachment of proteins, both shear and viscosity were increased relative to physically adsorbed proteins. An increase in lateral density of protein-binding surface of the SAM components was shown to increase the shear/viscosity of the resultant film while an increase in distance from the electrode (through incorporation of lysine linkers) was shown to decrease the shear/viscosity while simultaneously increasing the wet mass gain of the films. Shear and viscosity may be indicative of both enzyme denaturation and increased lateral protein packing within the film structure hence it is assumed that less distortion occurs with the inclusion of linkers which allow for more optimal protein immobilization.     

Construction of viscoelastic biocompatible films via the layer-by-layer assembly of hyaluronan and phosphorylcholine-modified chitosan

Films of hyaluronan (HA) and a phosphorylcholine-modified chitosan (PC-CH) were constructed by the polyelectrolyte multilayer (PEM) deposition technique and their buildup in 0.15 M NaCl was followed by atomic force microscopy, surface plasmon resonance spectroscopy (SPR), and dissipative quartz crystal microbalance (QCM). The HA/PC-CH films were stable over a wide pH range (3.0-12.0), exhibiting a stronger resistance against alkaline conditions as compared to HA/CH films. The loss and storage moduli, G' and G", of the films throughout the growth of eight bilayer assemblies were derived from an impedance analysis of the QCM data recorded in situ. Both G' and G" values were one order of magnitude lower than the moduli of HA/CH films. The fluid gel-like characteristics of HA/PC-CH multilayers were attributed to their high water content (50 wt %), which was estimated by comparing the surface coverage values derived from SPR and QCM measurements. Given the versatility of the PEM methodology, HA/PC-CH films are attractive tools for developing biocompatible surface coatings of controlled mechanical properties.     

Solution structure of the Vts1 SAM domain in the presence of RNA

The yeast Vts1 SAM (sterile alpha motif) domain is a member of a new class of SAM domains that specifically bind RNA. To elucidate the structural basis for RNA binding, the solution structure of the Vts1 SAM domain, in the presence of a specific target RNA, has been solved by multidimensional heteronuclear NMR spectroscopy. The Vts1 SAM domain retains the "core" five-helix-bundle architecture of traditional SAM domains, but has additional short helices at N and C termini, comprising a small substructure that caps the core helices. The RNA-binding surface of Vts1, determined by chemical shift perturbation, maps near the ends of three of the core helices, in agreement with mutational data and the electrostatic properties of the molecule. These results provide a structural basis for the versatility of the SAM domain in protein and RNA-recognition.     

Requirement of homeobox gene STIMPY/WOX9 for Arabidopsis meristem growth and maintenance

Most organs of flowering plants develop postembryonically from groups of pluripotent cells called meristems [1]. The shoot apical meristem (SAM) is specified by two complementary pathways [2-4]. SHOOT MERISTEMLESS (STM; [5]) defines the entire SAM region [6]. WUSCHEL (WUS), on the other hand, functions in a more restricted set of cells to promote stem-cell fate and is regulated by the CLAVATA genes in a negative feedback loop [7-10]. In contrast, little is known about how the growth of the SAM, which increases in size during vegetative development [11], is regulated. We have characterized STIMPY (STIP; also called WOX9 [12]), a homeobox gene required for the growth of the vegetative SAM, in part by positively regulating WUS expression. In addition, STIP is required in several other aerial organs and the root. What sets STIP apart from STM and WUS is that stip mutants can be fully rescued by stimulating the entry into the cell cycle with sucrose. Therefore, STIP is likely to act in all these tissues by maintaining cell division and preventing premature differentiation. Taken together, our findings suggest that STIP identifies a new genetic pathway integrating developmental signals with cell-cycle control.     

Immunoassay of <Emphasis Type="Italic">S</Emphasis>-adenosylmethionine and <Emphasis Type="Italic">S</Emphasis>-adenosylhomocysteine: the methylation index as a biomarker for disease and health status

Background

S-Adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are relevant to a variety of diseases. Previous reports that quantified SAM and SAH were based on HPLC or LC–MS/MS. No antibody against SAM has been generated, and the antibody against SAH cannot be used with blood samples. Immunoassays have not been used to measure SAM and SAH. In this study, ELISA was used to measure blood SAM and SAH levels.

Results

Specific antibodies against SAM were produced for the first time using a stable analog as the antigen. The monoclonal antibodies against SAM and SAH were characterized. No cross-reactivity was detected for the analyzed analogs. For the anti-SAM antibodies, the ELISA sensitivity was ~2 nM, and the affinity was 7.29 × 10¹⁰ L/mol. For the anti-SAH antibodies, the sensitivity was ~15 nM, and the affinity was 2.79 × 10⁸ L/mol. Using high-quality antibodies against SAM and SAH, immunoassays for the detection of SAM and SAH levels in blood and tissue samples were developed. Clinical investigations using immunoassays to measure SAM, SAH and the methylation index (MI) in normal and diseased samples indicated that (1) the SAM level is age and gender dependent; (2) the SAM level is associated with the severity of liver diseases, inflammatory reactions and other diseases; and (3) the methylation index (MI) is significantly reduced in many diseases and may serve as a screening biomarker to identify potentially unfavorable health conditions.

Conclusion

It is possible to generate antibodies against active small biomolecules with weak immunogenicity, such as SAM and SAH, using traditional hybridoma technology. The antigens and antibodies described here will contribute to the development of immunoassays to measure SAM, SAH and related molecules. These assays enable the MI to be measured specifically, accurately, easily and quickly without costly equipment. This preliminary study indicates that the MI could be an effective indicator of general health, except under conditions that may alter the value of the MI, such as special diets and medications.

     

Initiation of shoot apical meristem in rice: characterization of four SHOOTLESS genes.

The regulatory mechanism of shoot apical meristem (SAM) initiation is an important subject in developmental plant biology. We characterized nine recessive mutations derived from four independent loci (SHL1-SHL4) causing the deletion of the SAM. Radicles were produced in these mutant embryos. Concomitant with the loss of SAM, two embryo-specific organs, coleoptile and epiblast, were lost, but the scutellum was formed normally. Therefore, differentiation of radicle and scutellum is regulated independently of SAM, but that of coleoptile and epiblast may depend on SAM. Regeneration experiments using adventitious shoots from the scutellum-derived calli showed that no adventitious shoots were regenerated in any shl mutant. However, small adventitious leaves were observed in both mutant and wild-type calli, but they soon became necrotic and showed no extensive growth. Thus, leaf primordia can initiate in the absence of SAM, but their extensive growth requires the SAM. An in situ hybridization experiment using a rice homeobox gene, OSH1, as a probe revealed that shl1 and shl2 modified the expression domain of OSH1, but normal expression of OSH1 was observed in shl3 and shl4 embryos. Accordingly, SHL1 and SHL2 function upstream of OSH1, and SHL3 and SHL4 downstream or independently of OSH1. These shl mutants are useful for elucidating the genetic program driving SAM initiation and for unraveling the interrelationships among various organs in grass embryos.     

植物茎尖分生组织分化调控机制研究进展

茎尖分生组织是位于植物顶端具有持续分化能力的组织,通过细胞分裂、分化产生茎、叶和花等器官,形成植株地上部分。茎尖分生组织在分化过程中受外界环境因素、内源激素水平和分子调控等影响,表现出明显变化。该文综合国内外近年来有关茎尖分生组织分化调控的研究进展,从茎尖分生组织的形态结构和环境影响因素,以及激素调控和分子调控等方面,对茎尖分生组织分化活动的研究进行综述,并对目前研究现状存在问题及未来研究方向进行了分析和展望。     

On the mechanism of air ventilaton in anabantoids (Pisces: Teleostei)

Summary Air ventilation in most Anabantoid species is diphasic, consisting of exhalation and inhalation. Exhalation is the release of air from the accessory breathing organs (suprabranchial chambers) through the mouth either into the water near the surface (e.g.,Ctenopoma) or directly into the atmosphere (e.g.,Osphronemus goramy). Inhalation, i.e., taking in fresh air through the mouth at the surface, immediately follows exhalation. X-ray films show (Figs. 5 and 6) that evacuation of the suprabranchial chambers during exhalation is total or nearly total. This, together with the fact that these chambers can contract at most to a very small extent, led to the conclusion that gas is replaced by water entering the chambers during exhalation and that this water is replaced by fresh air during inhalation. Further analysis of films, including conventional films showing the behavior of the opercular apparatus during air ventilation (Fig. 7), leads to a theory of a double-pumping mechanism responsible for air ventilation. This mechanism consists of the buccal apparatus and the opercular apparatus. It is suggested that both of these structures are able to act as both suction and pressure pumps, and thus air ventilation may be explained as the result of alternating activity of these two pumps.In the monophasic air ventilation characteristic of (adult)Anabas testudineus, there is no exhalation phase comparable to that of other Anabantoids. Therefore, no water enters the suprabranchial chambers, which remain filled with gas during the whole ventilation process (Fig. 10). Ventilation is limited to one phase comparable to inhalation in other Anabantoids.The structure of the accessory breathing organs (Fig. 1) and its progressive complication with growth (Fig. 4) were studied inOsphronemus goramy. The arrangement of the labyrinthine plates is in accordance with the requirements of transport of water and gas through the suprabranchial chambers. One plate (the inner plate, Fig. 1) separates these chambers into atrium, ventro-caudal, and dorso-caudal compartments, each with its own opening (valve). This organization seems essential for the transport of gas and water through the suprabranchial chambers and ensures that during exhalation, water flows into the chambers from above, so that while water is filling these chambers displaced gas can be sucked through the deep-lying pharyngeal openings into the expanding buccal cavity.Supported by the Deutsche Forschungsgemeinschaft     

Role of S-adenosyl-L-methionine in potentiating cadmium mobilization by diethylenetriamine penta acetic acid in mice

The beneficial effects of S-adenosyl-L-methionine (SAM) in potentiating the mobilization of cadmium by cadmium trisodium diethylenetriamine penta acetic acid (DTPA) from the major target organs and restoration of depleted tissue glutathione (GSH), zinc and copper concentration, were determined in cadmium-exposed mice. The results indicated a significant depletion of cadmium concentration from the blood in DTPA plus SAM treated animals compared with DTPA or SAM alone treated groups. The treatment with SAM alone was also effective in correcting the zinc and GSH concentrations. The results indicated few beneficial effects of concomitant SAM administration during chelation of cadmium with DTPA.     

Establishment of the embryonic shoot apical meristem in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

In higher plants, shoot organs such as leaves, branches, and flowers are generated from the shoot apical meristem (SAM), a small group of undifferentiated cells located at the tip of the shoot. The SAM maintains its pluripotency and simultaneously produces lateral organs at its periphery. The SAM arises during embryogenesis and its positioning requires axis-dependent embryo patterning and compartmentalization of the embryo apex. Here, we introduce major factors involved in these processes in Arabidopsis thaliana and discuss how the embryonic SAM is established.     

拟南芥茎顶端分生组织相关突变体的表型与结构分析

以拟南芥野生型(C24)和T-DNA插入诱发的突变体(155系)为材料,通过表型分析、组织切片、GUS基因表达的组织化学定位等研究方法对155系的形态结构和生长发育进行了较为细致的观察分析,结果发现:(1)T-DNA插入诱发的155系突变体植株矮化,叶片等器官体积减小,营养生长阶段延长,发育较C24缓慢;(2)同一时期155系的茎顶端分生组织面积较C24减小,顶端平坦,细胞层数减少,两侧叶原基基部之间的距离缩短,呈现出发育迟缓、从茎顶端分生组织向花分生组织转变延迟等特征;(3)GUS基因特异性地在155系茎顶端分生组织和维管组织中表达.结果表明,T-DNA诱捕基因可能在茎顶端分生组织中发挥作用,由于T-DNA的插入使该基因的功能受到了影响,进而影响了155系中茎顶端分生组织的发育模式,产生了155系的一系列表型改变.     

Selective enhancement of Raman or fluorescence spectra of biomolecules using specifically annealed thick gold films

Annealing of "thick" metal films deposited onto a smooth dielectric substrate leads to high-order self-organization of metal clusters on the film surface. This work presents the first experimental evidence that the "thick" gold film (TGF) may be specifically annealed to serve as a substrate for surface-enhanced fluorescence or surface-enhanced Raman scattering (SERS) spectroscopy of the same molecule. High-resolved SERS spectra of mitoxantrone (mitox) were recorded on the TGF annealed at 340 degrees C whereas no Raman enhancement but an increase of mitox fluorescence signal were detected on the TGF annealed at 240 degrees C. The mitox fluorescence was further enhanced by deposition of monolayers of pentanethiol or poly-L-lysine on the surface of annealed TGF. The maximal fluorescence enhancement factor per mitox molecule of approximately 50 that was obtained on the annealed TGF covered with poly-L-lysine makes the system promising for applications in immunofluorescence assays and in microspectrofluorescence analysis.     

Plant stem cells and their regulations in shoot apical meristems

Stem cells in plants, established during embryogenesis, are located in the centers of the shoot apical meristem (SAM) and the root apical meristem (RAM). Stem cells in SAM have a capacity to renew themselves and to produce new organs and tissues indefinitely. Although fully differentiated organs such as leaves do not contain stem cells, cells in such organs do have the capacity to re-establish new stem cells, especially under the induction of phytohormones in vitro. Cytokinin and auxin are critical in creating position signals in the SAM to maintain the stem cell organizing center and to position the new organ primordia, respectively. This review addresses the distinct features of plant stem cells and focuses on how stem cell renewal and differentiation are regulated in SAMs.